Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Basal cells of the human airways acquire mesenchymal traits in idiopathic pulmonary fibrosis and in culture.

doi: 10.1038/labinvest.2015.114

Figure Lengend Snippet: Figure 1 Basal cell reactivity, hyperplasia and upregulation of EMT markers adjacent to fibroblastic foci. Epithelium adjacent to fibroblastic foci shows increased reactivity and mesenchymal phenotype. Sparse staining of CK14 is seen in control samples, whereas CK14 is strongly expressed in epithelium overlying fibroblastic foci. P63, a restricted transcription factor for basal cells is expressed in epithelial cells located both basally and in the top layer indicating formation of metaplasia in IPF samples. E-cadherin is abundant in epithelium both in control and IPF samples. In addition, E-cadherin is present in cells within the foci. N-cadherin is expressed in lower basal epithelial cells in IPF samples whereas control samples are negative. Vimentin is expressed in both epithelium and mesenchyme in IPF samples. Bar 100 μm.

Article Snippet: Lentiviral transduction was performed as previously described.28 In short, plasmids containing scrambled hairpin (pLKO.1 shSCR; Addgene plasmid 17920)29 or shRNA against p63 (shp63alpha pLKO.1 puro; Addgene plasmid 19120)30 were used with packaging plasmids (psPAX2 and pMD2.G) (Addgene plasmids 12260 and 12259, respectively) to generate viral titer in HEK-293 T cells using Arrest-in (Open Biosystems).

Techniques: Staining, Control

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Basal cells of the human airways acquire mesenchymal traits in idiopathic pulmonary fibrosis and in culture.

doi: 10.1038/labinvest.2015.114

Figure Lengend Snippet: Figure 5 Knockdown (KD) of p63 abrogates UG-induced EMT of VA10 cells. p63 KD cells are unable to undergo UG-induced EMT. The expression of the mesenchymal markers N-cadherin and Thy-1 is abrogated in treated KD cells, whereas scrambled control cells show a division into two cellular sub-populations much like the mother cell line VA10. The expression of Vimentin can still be detected in treated KD cells, although to a lesser extent than in treated scrambled cells. Bars 100 μm.

Article Snippet: Lentiviral transduction was performed as previously described.28 In short, plasmids containing scrambled hairpin (pLKO.1 shSCR; Addgene plasmid 17920)29 or shRNA against p63 (shp63alpha pLKO.1 puro; Addgene plasmid 19120)30 were used with packaging plasmids (psPAX2 and pMD2.G) (Addgene plasmids 12260 and 12259, respectively) to generate viral titer in HEK-293 T cells using Arrest-in (Open Biosystems).

Techniques: Knockdown, Expressing, Control